Differentiation of glutamatergic neurons from mouse embryonic stem cells requires raptor S6K signaling
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Updated: May 8, 2026
Differentiation of Mouse Embryonic Stem Cells into Cortical Interneuron Precursors
Published on: December 3, 2017
Jen-Hua Chuang1, Li-Chu Tung, Yi Yin
1Department of Life Science, National Taiwan Normal University, Taipei 116,Taiwan.
Abstract:
Although the mammalian target of rapamycin complex 1 (mTORC1) functions as an important signaling complex in many cellular processes, the role of mTORC1 in neurons derived from embryonic stem cells (ESCs) has been less explored. Here, using a modified protocol to differentiate mouse ESCs (mESCs) into almost uniform glutamatergic neurons, we explored the importance of raptor/mTORC1 in the differentiation of mESCs. Raptor gene-trap mESCs, and raptor-knockdown mESCs formed smaller-sized embryonic bodies than the wild type and failed to undergo neuronal differentiation. Treatment with 1μM rapamycin starting at the point when neuronal precursors began to differentiate from mESCs caused the gradual loss of neurites, shrinkage of soma, and a decreased ratio of neurite length to cell number over 48 to 72h of treatment. This change was accompanied by activation of caspase-3 and S6 kinase (S6K), but not 4E-binding protein 1 (4EBP1). Knockdown of raptor during neuronal differentiation from mESCs also resulted in gradual loss of neurites and shrinkage of cell bodies. Loss of neurite density resulting from rapamycin treatment could be reversed by overexpression of S6K T389E. Taken together, these data demonstrate that raptor/mTORC1/S6K plays a critical role in the differentiation and survival of neurons derived from mESCs.
Insights
The mammalian target of rapamycin complex 1 (mTORC1) is crucial for neuronal differentiation from mouse embryonic stem cells (mESCs). Inhibiting mTORC1 impairs neuronal development and survival, highlighting its essential role in neuron formation.
Area of Science:
- Cell Biology
- Neuroscience
- Stem Cell Biology
Background:
- Mammalian target of rapamycin complex 1 (mTORC1) is a key regulator of cellular processes.
- Its specific role in neurons derived from embryonic stem cells (ESCs) remains underexplored.
Purpose of the Study:
- To investigate the function of raptor/mTORC1 in the differentiation of mouse ESCs (mESCs) into glutamatergic neurons.
- To elucidate the impact of mTORC1 signaling on neuronal development and survival.
Main Methods:
- Utilized a modified protocol for differentiating mESCs into glutamatergic neurons.
- Employed raptor gene-trap and raptor-knockdown mESCs to assess mTORC1 function.
- Administered rapamycin to inhibit mTORC1 activity during neuronal differentiation.
- Analyzed cellular morphology, neurite outgrowth, and apoptosis markers (caspase-3).
- Investigated the role of S6 kinase (S6K) in mTORC1-mediated effects.
Main Results:
- Raptor deficiency in mESCs resulted in smaller embryonic bodies and impaired neuronal differentiation.
- Rapamycin treatment led to neurite loss, soma shrinkage, and reduced neurite length.
- These effects were associated with caspase-3 activation and S6K activation, but not 4E-binding protein 1 (4EBP1).
- Raptor knockdown mirrored rapamycin's effects on neurite loss and cell body shrinkage.
- Overexpression of a constitutively active S6K mutant reversed rapamycin-induced loss of neurite density.
Conclusions:
- Raptor/mTORC1 signaling is critical for the differentiation of mESCs into neurons.
- The mTORC1/S6K pathway plays a vital role in maintaining neuronal structure and survival.
- Targeting mTORC1 may offer therapeutic strategies for neurological disorders affecting neuronal development and maintenance.
